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Evaluation of the Bone Regeneration Effect of Recombinant Human Bone Morphogenic Protein-2 on Subperiosteal Bone Graft in the Rat Calvarial Model

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Evaluation of the Bone Regeneration Effect of Recombinant Human Bone Morphogenic Protein-2 on Subperiosteal Bone Graft in the Rat Calvarial Model

Eunhye Jang et al. Materials (Basel).

Abstract

The aim of this study was to evaluate the bone regeneration effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on a subperiosteal bone graft in a rat model. A subperiosteal space was made on the rat calvarium, and anorganic bovine bone (ABB), ABB/low bone morphogenetic protein (BMP) (5 µg), and ABB/high BMP (50 µg) were grafted as subperiosteal bone grafts. The new bone formation parameters of bone volume (BV), bone mineral density (BMD), trabecular thickness (TbTh), and trabecular spacing (TbSp) were evaluated by microcomputed tomography (µ-CT), and a histomorphometric analysis was performed to evaluate the new bone formation area. The expression of osteogenic markers, such as bone sialoprotein (BSP) and osteocalcin, were evaluated by immunohistochemistry (IHC). The ABB/high BMP group showed significantly higher BV than the ABB/low BMP (p = 0.004) and control groups (p = 0.000) and higher TbTh than the control group (p = 0.000). The ABB/low BMP group showed significantly higher BV, BMD, and TbTh than the control group (p = 0.002, 0.042, and 0.000, respectively). The histomorphometry showed significantly higher bone formation in the ABB/low and high BMP groups than in the control group (p = 0.000). IHC showed a high expression of BSP and osteocalcin in the ABB/low and high BMP groups. Subperiosteal bone grafts with ABB and rhBMP-2 have not been studied. In our study, we confirmed that rhBMP-2 contributes to new bone formation in a subperiosteal bone graft with ABB.

Keywords: bone morphogenetic protein; bone regeneration; bone sialoprotein; osteocalcin; subperiosteal bone graft.

Conflict of interest statement

The authors declare no conflict of interests.

Figures

Figure 1
Figure 1
Surgical procedure of the subperiosteal bone graft. A horizontal step incision was made on the posterior portion of calvarial bone, and subperiosteal dissection was performed. A subperiosteal pocket was made on the parietal bone of calvarium, and the anorganic bovine bone (ABB) alone and the ABB with 5 and 50 µg of recombinant human bone morphogenetic protein-2 (rhBMP-2) were grafted in the pocket.
Figure 2
Figure 2
The 3D reconstruction images of microcomputed tomography (μ-CT) in the (a) control, (b) ABB/low bone morphogenetic protein (BMP), and (c) ABB/high BMP group at 6 weeks after operation.
Figure 3
Figure 3
μ-CT analysis at 6 weeks after operation. (a) Bone volume, (b) bone mineral density, (c) trabecular thickness, and (d) trabecular spacing of the control (ABB: anorganic bovine bone), low BMP (ABB + 5 µg of rhBMP-2), and high BMP (ABB + 50 µg of rhBMP-2) group (** p < 0.05, *** p < 0.001).
Figure 3
Figure 3
μ-CT analysis at 6 weeks after operation. (a) Bone volume, (b) bone mineral density, (c) trabecular thickness, and (d) trabecular spacing of the control (ABB: anorganic bovine bone), low BMP (ABB + 5 µg of rhBMP-2), and high BMP (ABB + 50 µg of rhBMP-2) group (** p < 0.05, *** p < 0.001).
Figure 4
Figure 4
Histological images (hematoxylin and eosin staining) at 6 weeks after operation. (a,d) Control group, (b,e) ABB/low BMP, and (c,f) ABB/high BMP. (df) show high magnification of (ac), respectively. (ac): original magnification 40×, bar = 500 µm. (df): original magnification 100×, bar = 200 µm. ABB: anorganic bovine bone, NB: new bone.
Figure 5
Figure 5
Immunohistochemical staining of bone sialoprotein (BSP; ac) and osteocalcin (df) at 6 weeks after operation. (a,d) Control group, (b,e) ABB/low BMP, and (c,f) ABB/high BMP. A high expression of BSP and osteocalcin (black arrows) was observed in the new bone matrix of the ABB/low and high BMP groups (original magnification 100×, bar = 200 µm). ABB: anorganic bovine bone, NB: new bone.

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